【摘要】 目的研究去血清损伤中PKC信号通路的改变,同时阐明神经节苷脂对大鼠嗜铬细胞瘤细胞株(PC12)细胞去血清损伤的保护作用及其作用机制。方法采用去血清损伤24h作为损伤模型,MTT测定细胞存活率,Hoechst 33258/PI观察细胞损伤后的死亡类型及坏死、凋亡的细胞数;Western blotting检测PCl2细胞在损伤后细胞浆和细胞膜PKC蛋白的变化。结果去血清损伤时间依赖性对PC12细胞有损伤,多数细胞呈凋亡征象,而神经节苷脂对去血清损伤有明显的保护作用;去血清损伤时,PKC信号通路被活化,主要表现为PC12细胞胞浆中的PKC蛋白减少,胞膜上的PKC蛋白逐渐增加,实现PKC蛋白转位;而神经节苷脂可以抑制这种转位,从而起到保护作用。结论神经节苷脂对去血清损伤有保护作用,其保护作用部分是由于神经节苷脂抑制PKC信号通路的活化。更多还原

【Abstract】 Objective To determinted whether GM1 had a protective effect on injury induced by serum-deprivation and the possible mechanism in PC12 cells. Methods The viability of PC12 cells was quantified by MTT after serum-deprivation.The number of apoptotic cells and necrotic cells were determined by Hoechst 33258/PI staining.And the change of PKC protein expression on PC12 cells’ membrane and cytosols was detected by Western blotting. Results 1.The viability of PC12 cells decreased after serum-deprivation and the serum-deprivation for 24 hours was chosen as an injury model in this research.Most of the PC12 cells presented apoptosis 24 hours after serum-deprivation.In addition,the PC12 cells’ cytosols PKC protein decreased,while the PC12 cells’ membrane PKC protein increased significantly,and this result suggested PKC’s translocation to membrane and its activation.2.The viability of PC12 cells preincubated with GM1 in high concentrations(10,1,0.1μmol/L) increased significantly and GM1 protected PC12 cells from apoptosis after serum-deprived injury.GM1 reduced the damage of serum-deprivation on PC12 cells and inhibited PKC protein translocation after injury.3.The repair function of GM1 was effective to neuronal resume after serum-deprived injury.Conclusion Neuroprotective effects of GM1 on serum-deprived injury may be partly mediated through the regulation of PKC pathways and it is helpful for the recovery after injury.更多还原